Amphetamine In Blood Research Articles

Nystagmus among suspected amphetamine impaired drivers

Clinical signs of drug use can be helpful to identify which drug has been consumed. Amphetamine intake has traditionally not been considered to cause nystagmus. The aim of this study was to explore whether there is a relationship between amphetamine use and nystagmus in a population of apprehended drivers in a naturalistic setting. We evaluated drivers suspected of drug-impaired driving where blood samples were collected and a clinical test of impairment (CTI) was performed. Evaluation of nystagmus is one of the CTI subtests. The samples were analysed for alcohol and psychoactive drugs. Cases with a nystagmus test were recorded and amphetamine-only cases were compared with alcohol-only cases and with cases where alcohol or drugs were not detected, respectively. Samples from 507 amphetamine-only cases were compared to 485 alcohol-only cases and 205 drug-negative cases. The median blood amphetamine concentration was 0.37 mg/L and the median alcohol concentration was 1.57 g/kg. The proportion of cases with nystagmus was similar in amphetamine-only cases (21%) and drug-negative controls (25%), p = 0.273, but higher in alcohol-only cases (53%), p < 0.001. No association was found between the blood amphetamine concentration and degree of nystagmus (Spearman's ρ = 0.008, p = 0.860), whereas an association between blood alcohol concentration and degree of nystagmus was demonstrated (ρ = 0.249, p < 0.001). In conclusion, our study did not find that apprehended drivers using amphetamine had more frequently nystagmus than a control group that tested negative for alcohol and drugs, even at high amphetamine concentrations in blood. Hence, nystagmus should not be considered a tool for identifying amphetamine-induced impairment in drivers.

Nystagmus among amphetamine-impaired drivers

Clinical signs of drug use can be helpful to identify which drug has been used. Amphetamine intake has traditionally not been considered to cause nystagmus; however, there is a lack of evidence for this claim. The aim of this study was to find a relationship between amphetamine use and nystagmus in a population of apprehended drivers. We evaluated cases of drivers suspected of drugged driving where blood samples were collected and a clinical test of impairment (CTI) was performed. Evaluation of nystagmus is one of the CTI subtests. The samples were analysed for alcohol and psychoactive drugs. Cases with nystagmus were recorded and amphetamine-only cases were compared with alcohol-only cases and with cases where alcohol or drugs were not detected, respectively. Samples from 509 amphetamine-only cases were compared to 519 alcohol-only cases and 205 drug negative cases. The mean blood amphetamine concentration was 0.533 mg/L (range 0.027–3.650 mg/L) and the mean alcohol concentration was 1.57 g/kg (range 0.05–3.97 g/kg). The number of cases with nystagmus was similar in amphetamine-only cases (21%) and drug-negative controls (25%), P = 0.273. The number of cases with nystagmus was higher in alcohol-only cases (53%), than in amphetamine-only cases (21%), P < 0.001, and drug-negative controls (25%), P < 0.001. No association was found between the blood amphetamine concentration and nystagmus, whereas an association between blood alcohol concentration and nystagmus was demonstrated (Pearson's r = 0.252, P = 0.01). In our study, we did not find that apprehended drivers using amphetamine had more frequently nystagmus than a control group that tested negative for alcohol and drugs. One implication is that nystagmus is not a sensitive test to identify amphetamine-induced impairment.

A novel screening method for 64 new psychoactive substances and 5 amphetamines in blood by LC–MS/MS and application to real cases

Identification and quantification of new psychoactive substances (NPS), both in biological and non-biological samples, represent a hard challenge for forensic toxicologists. NPS are increasingly emerging on illegal drug market. Many cases of co-consumption of NPS and other substances have also been reported. Hence, the development of analytical methods aiming at the detection of a broad-spectrum of compounds (NPS and “traditional” drugs) could be helpful. In this paper, a fully validated screening method in blood for the simultaneous detection of 69 substances, including 64 NPS (28 synthetic cannabinoids, 19 synthetic cathinones, 5 phenethylamines, 3 indanes, 2 piperazines, 2 tryptamines, 2 phencyclidine, methoxetamine, ketamine and its metabolite) and 5 amphetamines (amphetamine, methamphetamine, MDMA, MDA, 3,4-methylenedioxy-N-ethylamphetamine − MDEA-) by a dynamic multiple reaction monitoring analysis through liquid chromatography − tandem mass spectrometry (LC–MS/MS) is described. This method is very fast, easy to perform and cheap as it only requires the deproteinization of 200μL of blood sample with acetonitrile. The chromatographic separation is achieved with a C18 column. The analysis is very sensitive, with limits of quantification ranging from 0.1 to 0.5ng/mL. The method is linear from 1 to 100ng/mL and the coefficient of determination (R2) was always above 0.9900. Precision and accuracy were acceptable at any quality control level and recovery efficiency range was 72–110%. Matrix effects did not negatively affect the analytical sensitivity.This method was successfully applied to three real cases, allowing identification and quantification of: mephedrone and methamphetamine (post-mortem); ketamine, MDMA and MDA (post-mortem); AB-FUBINACA (ante-mortem).

High prevalence of previous arrests for illicit drug use and/or impaired driving among drivers killed in motor vehicle crashes in Sweden with amphetamine in blood at autopsy

BackgroundAmphetamine, and to a lesser extent the secondary amine methamphetamine, are major recreational drugs of abuse in Sweden. These central stimulant amines are identified in blood from roughly 50% of people arrested for driving under the influence of drugs (DUID). However, much less information is available about the presence of amphetamine in blood of drivers killed in road-traffic crashes. MethodsThis retrospective 10-year study (2001–2010) used a forensic toxicology database (TOXBASE) to retrieve information about road-traffic crashes when the driver had amphetamine and/or methamphetamine in autopsy blood. Forensic toxicology results were available from over 95% of all drivers killed on Swedish roads during this 10-year period. ResultsAmphetamine was present in the blood of 106 drivers (3.9%) either alone or together with other psychoactive substances (e.g. alcohol, cannabis, diazepam, alprazolam, etc.). The vast majority of fatalities were male (95%) with a mean age (±standard deviation) of 37±11.4 years (range 16–67 years). The mean (median) and highest concentrations of amphetamine in femoral blood were 1.36mg/L (1.0mg/L) and 6.74mg/L, respectively. Many of the victims (75%) had been arrested previously for use of illicit drugs or DUID. The median number of previous arrests was 4 (range 0–83) and amphetamine or methamphetamine were among the drugs identified in blood samples from 89% of cases (0–100%). ConclusionThe high prevalence of repeat DUID offending and/or use of illicit drugs among the drivers killed in road-traffic crashes suggests that an early intervention and treatment for stimulant abuse might have been more beneficial than conventional punishments for such drug-related crimes.

MALDI–TOF mass spectrometric determination of four amphetamines in blood

A rapid and sensitive detection method using matrix-assisted laser desorption ionization time-of-flight mass spectrometry was developed for the analysis of amphetamine (A), methamphetamine (MA), 3,4-methylenedioxyamphetamine (MDA), and 3,4-methylenedioxymethamphetamine (MDMA). In this method, α-cyano-4-hydroxy cinnamic acid was used as the matrix to assist the ionization of amphetamines. The MS spectra of these amphetamines showed protonated molecules [M + H]+ and fragment ions with comparable or higher intensities. The quantifications of A and MA were performed using A-d 7 and MA-d 5 as the internal standard, respectively, and those of MDA and MDMA were performed using MDMA-d 5 as the internal standard. The limit of detection and the quantification range using 20 μl of blood were about 10 ng/ml and 30–500 ng/ml for A and MDA, respectively, and about 1 ng/ml and 3–50 ng/ml for MA and MDMA, respectively. In two cases of poisoning in which MA was abused, the levels of A in blood and urine were from 0.050 to 3.24 μg/ml and the levels of MA were from 0.231 to 25.1 μg/ml.

4-Methyl-amphetamine: a health threat for recreational amphetamine users

4-Methylamphetamine (4-MA) was originally developed as an appetite suppressant, but development was halted due to side effects. It has recently resurfaced as a new psychoactive substance in Europe, and is mostly found together with amphetamine. Around 11.5% of tested Dutch speed samples were positive for 4-MA. In Belgium, 4-MA was also found in speed samples. In 2011 and 2012, several fatal incidents after amphetamine use were observed in Belgium, the United Kingdom and The Netherlands. In all victims, toxicological analysis confirmed the presence of 4-MA, in addition to amphetamine. The observed blood amphetamine levels were too low to be fatal. Contrary to amphetamine, which displays noradrenergic and dopaminergic activity, 4-MA also shows serotonergic activity, which may contribute to the observed toxicity. Other mechanisms of toxicity are put forward in this paper as well. To conclude, the observed toxicity is most likely the result of the combined dopaminergic activity of amphetamine and the serotonergic activity of 4-MA. In addition, the presence of 4-MA may have dampened the psychoactive effects of amphetamine by attenuation of the amphetamine-induced dopamine release, potentially inclining users to ingest higher doses of contaminated speed. Also, slower metabolism of 4-MA and its MAO-inhibiting properties can also contribute to the unusual high toxicity of 4-MA.

Amphetamine Abuse in Sweden

Amphetamine is a major drug of abuse in Sweden and in the other Nordic countries. The demographics of amphetamine abusers in Sweden and the concentrations of this stimulant in blood are reported for 10 years of forensic blood samples (2001-2010). Using a forensic toxicology database (TOXBASE), we studied 1183 amphetamine-related deaths, 20,452 users of illicit drugs, and 47,366 people arrested for driving under the influence of drugs (DUID). Most amphetamine abusers were male (82%-87%), and their average age was 33 to 39 years with males being 2 to 3 years older than females (P < 0.001). Mean (median) concentrations of amphetamine in blood were 1.25 (0.40) mg/L in autopsy cases, 0.61 (0.40) mg/L in users of illicit drugs, and 0.76 (0.58) mg/L in DUID suspects. Median concentration in DUID suspects was significantly higher than in the other forensic materials (P < 0.001). Women also had higher median concentrations of amphetamine in blood than male abusers of this central stimulant (P < 0.001). The major coingested drugs were benzodiazepines (41%), cannabis (26%), opiates (21%), and alcohol (18%) in autopsy cases. Polydrug use was less common in DUID suspects and users of illicit drugs, although benzodiazepines (13%), tetrahydrocannabinol (12%), and opiates (5%) were often identified along with amphetamine. Because median concentration of amphetamine was higher in living subjects (DUID suspects) compared with amphetamine-related deaths, this points toward toxicity of coingested drugs or adverse drug-drug interaction as being responsible for death.

Concentration Ratios of Methamphetamine to Amphetamine in Blood Can Help to Distinguish Use of Methamphetamine from Various Mixtures of the Two Stimulants

Using a forensic toxicology database, the authors investigated cases of driving under the influence of drugs (DUID) if methamphetamine (MA) was identified in the blood samples (N = 9,310). The concentrations of MA and amphetamine (AM) in blood were determined after liquid-liquid extraction by gas chromatography-mass spectrometry at limits of quantitation of 0.03 mg/L for both stimulants. In 814 cases, AM was negative in blood and MA was positive at mean (median) and highest concentrations of 0.19 mg/L (0.11 mg/L) and 3.4 mg/L, respectively. Both amines were present in blood in 8,496 cases at concentrations of 0.54 mg/L (0.35 mg/L) and 10.4 mg/L for AM and 0.41 mg/L (0.22 mg/L) and 5.6 mg/L for MA. However, the correlation between AM and MA was low and insignificant (r = -0.13) in the whole material. The coefficient of correlation increased to r = 0.41 (P < 0.001) when the MA/AM concentration ratio was >1. When MA/AM ratios were selected at intervals of 1.0 (e.g., >3.0 and <4.0 up to >9.0 and <10.0), the correlation between AM and MA was r = 0.99 (P < 0.001). Such cases represent the use of MA without contamination from AM, and the mean (median) and highest concentrations of this secondary amine in blood of DUID suspects were 0.72 mg/L (0.56 mg/L) and 4.2 mg/L, respectively.

Estimating equivalent cutoff thresholds for drugs in blood and oral fluid using prevalence regression: A study of tetrahydrocannabinol and amphetamine

To validate a method for determining equivalent drug cutoff concentrations for tetrahydrocannabinol and amphetamine in blood and oral fluid, which ensures that the drug prevalence in samples of blood and oral fluid taken simultaneously is equal. A method using regression analysis of drug concentrations for defined percentiles in blood and oral fluid was developed. The accuracy and precision of this technique was investigated. As study populations, 311 cannabis users and 197 amphetamine users from the Rosita-2 Project were used. A total of 80 paired oral fluid and blood concentrations were needed to determine accurate regression formulae. When using the formulae to calculate drug cutoff concentrations in oral fluid corresponding to 2.0, 4.0, 6.0, 8.0 and 10.0 ng/ml tetrahydrocannabinol in blood and 200, 400, 600, 800 and 1000 ng/ml amphetamine in blood, the accuracy was better than 100 ± 20% compared to actual prevalence in blood with precision better than ± 20%. Prevalence regression may be a useful tool in estimating equivalent cutoff concentrations in blood and oral fluid.

High-Throughput Analysis of Amphetamines in Blood and Urine with Online Solid-Phase Extraction-Liquid Chromatography—Tandem Mass Spectrometry

An automated online solid-phase extraction-liquid chromatography-tandem mass spectrometry (SPE-LC-MS-MS) method for the analysis of amphetamines in blood and urine was developed and validated. Chromatographic separation was achieved on a Nucleodur Sphinx RP column with an LC gradient (a mixture of 10 mM ammonium formate buffer and acetonitrile), ensuring the elution of amphetamine, methamphetamine, MDMA, MDA, MDEA, PMA, and ephedrine within 11 min. The method was fully validated, according to international guidelines, using only 100 and 50 microL of blood and urine, respectively. The method showed an excellent intra- and interassay precision (relative standard deviation < 11.2% and bias < 13%) for two external quality control samples (QC) for both matrices and three and two 'in house' QCs for blood and urine, respectively. Responses were linear over the investigated range (r(2) > 0.99, 2.5-400 microg/L for blood and 25-1000 microg/L for urine). Limits of quantification were determined to be 2.5 and 25 microg/L for blood and urine, respectively. Limits of detection ranged from 0.05 to 0.5 microg/L for blood and 0.25 to 2.5 microg/L for urine, depending on the compound. Furthermore, the analytes and the processed samples were demonstrated to be stable (in the autosampler for at least 72 h and after three freeze/thaw cycles), and no disturbing matrix effects were observed for all compounds. Moreover, no carryover was observed after the analysis of high concentration samples (15,000 microg/L). The method was subsequently applied to authentic blood and urine samples obtained from forensic cases, which covered a broad range of concentrations. The validation results and actual sample analyses demonstrated that this method is rugged, precise, accurate, and well-suited for routine analysis as more than 72 samples are analyzed non-stop in 24 h with minimum sample handling. The combination of the high-throughput online SPE and the well-known sensitivity and selectivity assured by MS-MS resulted in the elimination of the bottleneck associated with the sample preparation requirements and provided increased sensitivity, accuracy, and precision.

Analysis of amphetamine, methamphetamine, methylenedioxyamphetamine and methylenedioxymethamphetamine in whole blood using in-matrix ethyl chloroformate derivatization and automated headspace solid-phase microextraction followed by GC-MS

The in-matrix alkyl chloroformate derivatization method for amphetamine, methamphetamine, methylenedioxyamphetamine (MDA), and methylene-dioxymethamphetamine (MDMA) was adapted for use with the whole blood matrix. This derivatization method was followed by automated headspace (HS)-solid-phase microextraction (SPME) and gas chromatography-mass spectrometry (GC-MS) analysis. The sensitivity of this method, expressed as limit of detection, was approximately 10 ng/ml for these analytes tested in the blood matrix, which was sufficient to detect toxic concentrations of amphetamines in blood. The limit of quantitation for target analytes ranged from 0.05 to 0.2 μg/ml. The intraday precision and accuracy studies generally showed satisfactory results for all target compounds except MDA, in which a larger variation was observed. The in-matrix ethyl chloroformate derivatization of amphetamine, methamphetamine, MDA, and MDMA for HS-SPME was tested in other matrices such as stomach fluid, bile, thoracic cavity fluid, vitreous humor, brain, liver, spleen, and skeletal muscle. As a result, stomach fluid, thoracic cavity fluid, and vitreous humor showed SPME efficiencies higher than that of whole blood; however, this method was not suitable for solid tissue matrices under the present conditions.

Driving Under the Influence of Central Stimulant Amines: Age and Gender Differences in Concentrations of Amphetamine, Methamphetamine, and Ecstasy in Blood

A zero-tolerance law for driving under the influence of drugs (DUID) was introduced in Sweden in 1999. This change in legislation has led to a 12-fold increase in the number of blood samples sent by the police for toxicological analysis. Here we report the age and gender of offenders, along with the concentrations of amphetamine, methamphetamine, and ecstasy (3,4-methylenedioxymeth-amphetamine) in blood samples analyzed since the institution of the new legislation. A forensic toxicology database (TOXBASE) was used to identify cases of DUID in which central stimulant amines were verified in blood during a 5-year period (2000-2004). Amphetamine was present in 15,898 of 26,556 cases of DUID (60%) either alone or together with other licit or illicit drugs. In 6,094 cases, amphetamine was the only psychoactive substance in blood at mean (median) and highest concentrations of 1.01 mg/L (0.80 mg/L) and 11.9 mg/L, respectively. The users of amphetamine were mainly men (85% vs 15% women; p < .001), and men tended to be a few years older than the women; the mean (SD) age for men was 37 (9.2) years and for women it was 35 (8.1) years (p < .001). In 644 cases, amphetamine and methamphetamine were present in blood samples at mean (median) concentrations of 0.85 mg/L (0.60 mg/L) and 0.34 mg/L (0.20 mg/L), respectively (p < .001). The mean (median) and highest concentrations of ecstasy in 493 DUID offenders were 0.23 mg/L (0.10 mg/L) and 3.5 mg/L, respectively. The mean age of ecstasy users was 26 (7.2) years, which was about 10 years younger than those using amphetamine (p < .001). The high prevalence of amphetamines in blood of apprehended drivers in Sweden verifies widespread use of these stimulants as recreational drugs. The findings from this study suggest that a zero-tolerance DUID law has not deterred offenders, which suggests that more attention should be given to the underlying substance-abuse problem instead of conventional penalties such as monetary fines and/or imprisonment.

High re-arrest rates among drug-impaired drivers despite zero-tolerance legislation

Background A zero-tolerance law for driving under the influence of drugs (DUID) in Sweden led to a 10-fold increase in the number of cases submitted by the police for toxicological analysis. The statutory blood–alcohol concentration (BAC) limit for driving is 0.2 mg/g (∼0.02 g%). Methods An in-house database (TOXBASE) was used to investigate re-arrests for impaired driving over 4 years (2001–2004), which comprised 36,799 cases. The age, gender, re-arrest rate of the offenders and the concentrations of ethanol and amphetamine in blood samples were evaluated. Results We found that 44% of individuals ( N = 16,277) re-offended 3.2 times on average (range 1–23 arrests). Between 85 and 89% of first-time offenders were men and there was also a male dominance among the recidivists (88–93%). The mean age of drunken drivers was ∼40 years compared with ∼35 years for driving under the influence of amphetamine, which was the drug identified in 50–60% of DUID cases, either alone or together with other licit or illicit drugs. The median BAC was 1.5 mg/g (∼0.15 g%), which suggests a dominance of heavy drinkers. The median BAC was even higher in recidivists (1.6–1.7 mg/g). The median concentration of amphetamine in blood was 1.0 mg/L in recidivists compared with 0.5 mg/L in the first-time offenders. About 14% of drunken drivers re-offended 1–10 times compared with 68% of DUID suspects, who were re-arrested 1–23 times. People with only a scheduled prescription drug in blood were re-arrested much less frequently (∼17%) compared with those taking illicit drugs (68%). Conclusions The appreciable increase in number of arrests for DUID after a zero-tolerance law might reflect a heightened enthusiasm by the police authorities armed with knowledge that a prosecution is easier to obtain. Zero-tolerance laws do not deter people from impaired driving judging by the high re-arrest rates. During the sentencing of hardcore offenders, the courts should give more consideration to the underlying substance abuse problem.

Age‐ and gender‐related differences in blood amphetamine concentrations in apprehended drivers: lack of association with clinical evidence of impairment

New legislation aimed at combating driving under the influence of drugs (DUID) in Sweden stipulated zero-concentrations in blood for scheduled substances. DUID suspects (n = 300), with amphetamine as the only psychoactive drug identified in blood, were investigated in relation to age and gender. In a smaller retrospective sample (n = 70) the relationship between clinical tests of impairment were compared with the concentration of amphetamine in blood. All forensic blood samples were subjected to a broad toxicological screening analysis by immunoassay methods [enzyme multiplied immunoassay technique/cloned enzyme donor immunoassay (EMIT/CEDIA)] and positive results were verified by gas chromatography-mass spectrometry (GC-MS). The limit of quantitation (LOQ) for determination of amphetamine in blood was 0.03 mg/l. People suspected of being under the influence of drugs were examined by a physician who asked various questions about state of health and use of drugs and also administered simple psychomotor and cognitive tests of impairment. After conducting these tests the physician concluded whether the suspects were not impaired, slightly, moderately or highly impaired by drugs other than alcohol. Among 300 DUID suspects with amphetamine in blood there were 246 men (82%) and 54 women (18%). Mean age (+/- SD) of the men was 37.1 +/- 8.7 years compared with 35.5 +/- 7.1 years for the women (P > 0.05). The frequency distribution of blood amphetamine concentration was positively skewed with mean, median and highest values of 1.0 mg/l, 0.9 mg/l and 7.1 mg/l, respectively. The mean concentrations were slightly higher in the women 1.11 mg/l (median 1.0 mg/l) compared with 0.97 mg/l (median 0.8 mg/l) in the men (P > 0.05). There was a weak but statistically significant correlation between the person's age and the concentration of amphetamine in blood (r = 0.18, P < 0.05). The results of clinical tests of impairment showed no relationship with the concentration of amphetamine in blood according to analysis of variance (P > 0.05). The lack of association between degree of drug influence and the concentration of amphetamine in blood speaks against the notion of introducing concentration per se limits or graded penalties depending on the blood-concentration of this stimulant. Zero-concentration limits or LOQ-limits are a much more pragmatic way to enforce DUID legislation.

Impairment related to blood amphetamine and/or methamphetamine concentrations in suspected drugged drivers

Experimental studies have investigated effects of low oral doses of amphetamine and methamphetamine on psychomotor functions, while less work has been done on effects of high doses taken by abusers in real-life settings. There are indications that intake of high doses may impair traffic related skills, and that abuse of amphetamines may cause hypersomnolence at the end-of-binge. The present study aimed at investigating the concentration–effect relationship between blood amphetamines concentrations and impairment in a population of real-life users. Eight hundred and seventy-eight cases with amphetamine or methamphetamine as the only drugs present in the blood samples were selected from the impaired driver registry at The Norwegian Institute of Public Health. In each case the police physician had concluded on whether the driver was impaired or not. 27% of the drivers were judged as not impaired, while 73% were judged as impaired. There was a positive relationship between blood amphetamines concentrations and impairment. The relationship reached a ceiling at blood amphetamines concentrations of 0.27–0.53 mg/l. Younger drivers were more often judged impaired than older drivers at similar concentrations. Despite the performance enhancing qualities of amphetamines demonstrated in some low dose laboratory experiments; this study revealed a positive relationship between blood amphetamines concentration and traffic related impairment.

Abnormally High Concentrations of Amphetamine in Blood of Impaired Drivers

We present a case series (N = 46) of individuals apprehended in Sweden for driving under the influence of drugs (DUID). These cases were selected because the concentrations of amphetamine in blood were abnormally high (> 5.0 mg/L), the highest being 17 mg/L. In comparison, the median blood-amphetamine concentration in a population of DUID offenders (N = 6,613) was 0.70 mg/L. Among the DUID suspects with extremely high blood-amphetamine concentrations there were 38 men (83%) with mean age of 37.8 y (SD 6.8 y) and 8 women (17%) with a mean age of 34.1 y (SD 4.3 y). All had previously been registered in our database (mean 12 times, median 9 times) for drug-related offences, including DUID. The concentration of amphetamine in blood of female offenders was slightly higher than the concentration in male offenders (6.6 mg/L vs. 5.8 mg/L), although this difference was not statistically significant (p > 0.05). The drugs other than amphetamine most frequently encountered in the blood samples were tetrahydrocannabinol and benzodiazepines (diazepam and nordiazepam). The commonest signs of drug use reported by the arresting police officers were bloodshot and glazed (watery) eyes, restlessness, talkativeness, exaggerated reflexes and slurred speech. Unsteady gait and dilated pupils were observed in some but not all individuals. These very high concentrations of amphetamine were tolerated without any fatalities indicating a pronounced adaptation to the pharmacologic effects of this central stimulant. Anecdotal information indicated that those with the very highest concentrations of amphetamine in blood had swallowed the drug to prevent being apprehended in possession of an illicit substance.

Automated on-line dialysis and liquid chromatography of methylenedioxylated amphetamines in plasma and serum samples

An automated on-line dialysis coupled to a trace enrichment method has been developed for the separation and quantification of four methylenedioxylated amphetamines in serum and plasma, using liquid chromatography coupled to a fluorimetric detector. The on-line dialysis method was optimized and validated on fresh human serum and plasma samples. This sample preparation method allowed the quantification of methylenedioxylated amphetamines in serum or plasma, at concentrations as low as ca. 10 ng ml −1, with good repeatability, reproducibility and accuracy. The automated on-line dialysis method took less than 30 min. This method was applied to seven toxicological cases and results showed that the concentration of methylenedioxylated amphetamines in blood was in the range of 20–484 ng ml −1.

Rapid and simple determination of methamphetamine and amphetamine in blood by simultaneous extraction-derivatization

A method capable of extracting and derivatizing methamphetamine (MA) and amphetamine (AM) in blood simultaneously was developed. In this method ethyl acetate, triethylamine and pentafluorobenzyl bromide were added to blood, the extraction and pentafluorobenzyl derivatization of MA and AM were performed simultaneously by heating and stirring on a hot plate stirrer. Pentafluorobenzyl derivatives were determined by gas chromatography/chemical ionization-mass spectrometry (GC/CI-MS). Deuterium labeled MA and AM were used as internal standards. This method enabled an accurate and precise quantitative analysis from blood containing MA and AM. Excellent results were obtained for autopsy cases with possible MA.

Rapid analysis of amphetamines in blood using head space-solid phase microextraction and selected ion monitoring

A simple and rapid method for analysis of methamphetamine (MA) and amphetamine (AP) in blood was developed using head space-solid phase microextraction (HS-SPME) and gas chromatography-mass spectrometry/electron impact ionization-selected ion monitoring (GC-MS/EI-SIM). A vial containing a blood sample, sodium hydroxide, and pentadeuterated methamphetamine as an internal standard, was heated at 80 °C for 20 min. The extraction fiber of the SPME was exposed for 5 min in the head space of the vial. First, heptafluorobutyric anhydride solution was injected into the injection port of the GC-MS to make heptafluorobutyramide (HFB) derivatives of amphetamines, and compounds absorbed on the fiber were detached by exposing the fiber in the injection port. Straight calibration curves of MA and AP were obtained from 0.01 to 2 μg/g in blood, respectively. No interfering substances were found, and the time for analysis was 30 min for one sample.

Determination of methamphetamine and its metabolite amphetamine in biological fluids from 11 fatal cases

Gas chromatography with flame ionization detector (FID) along with mass spectrometry (GC/MS) were used for the screening and quantification of methamphetamine (MA) and its major metabolite, amphetamine (AM), in blood and urine in eleven fatal cases in which MA abuse was suspected. Postmortem blood MA varied from 0.5–30.2 μg/ml, while AM levels ranged from none detected (6 of 11 cases) to 4.8 μg/ml. Additionally, distribution studies were performed in three of these cases in which tissue samples were available for evaluation. Liver contained the highest concentration of MA among the tissue samples. In eight of the eleven cases, when no other direct cause of death was evident (i.e. 3 cases of traumatic death), either no blood AM was found or the ratio of MA/AM was 3.4 or greater. These data are consistent with acute MA use followed by death due to acute drug intoxication or by the occurrence of hypersensitivity and reverse tolerance seen in cases of chronic drug abuse.